6 10 Introduction: The application and use of criteria This chapter provides background information on the considerations which have led the Commission to propose microbiological criteria for some commodities and not others. It also indicates how the criteria should be interpreted and applied. A. FORMAT OF COMMODITY CHAPTERS In the first edition of this book foods were grouped on the basis of commodities (e.g., milk and milk products) or processes (e.g., frozen foods, dried foods). In this edition the commodities grouping used in Microbial Ecology of Foods, vol. 2 (ICMSF 1980), has been followed with two exceptions. These are formulated foods, comprising raw materials from several commodity groups, and low-acid canned products. Criteria for formulated foods will depend on conditions of manufacture, the types of raw materials used, the process, the intended distribution system, and shelf-life. Hence criteria are proposed only where a need has been demonstrated and such information is available (see Chapter 24, Formulated Foods). Sampling plans and microbiological tests are not relevant to the safety of shelf-stable canned foods and are therefore not proposed. Extensive cross-reference is made to the volume referred to above (ICMSF 1980) so that information pertaining to the need for criteria and the appropriate cases may readily be found. Each chapter discusses the microbiological hazards associated with a commodity group and, based on a consideration of their relevance, may propose criteria. General sampling procedures are dealt with in Chapter 9, but if a commodity requires special sampling procedures applications. 2nd Ed. International Commission on Microbiological Specifications for Foods.

7 128 Proposals for sampling and sample plans these are detailed within the chapter. For methods, reference is made to ICMSF, 1978, or to other appropriate sources. B. SELECTION AND APPLICATION OF CRITERIA Ideally the control of microorganisms in foods is at the point of production, processing, or preparation for consumption (see Chapter 7, Control at Source). However, for much food in international trade there is no knowledge of control at source or of the conditions used during processing and handling. Therefore, there remains a need for criteria to assess the acceptability of foods at port of entry. Before recommending a criterion for a product, the Commission decided that each product must meet the following conditions: 1 The product must be in international trade. 2 There must be good epidemiological evidence that the product, or product group, has been implicated in food-borne disease and/or may have an inadequate shelf-life if Good Commercial Practice (GCP) has not been followed. 3 There must be good evidence, or good reason to believe, that the application of a criterion will reduce the health risk in food and/ or effectively assess adherence to MCP. It is important to consider some of the difficulties in expecting a microbiological examination to portray the true microbiological condition of food. For example, a relatively few sample units may not accurately reflect the true microbiological status of a consignment, particularly if the consignment is composed of several batches of food. In this case, if only one of the batches is defective, sampling a portion of the consignment may not reveal the unacceptable part. The resulting decision, if applied to the whole consignment, would be quite unsatisfactory. The sampling plans proposed in this book should whenever possible be applied to identifiable lots (see definition of a lot, Chapter 3, Section A). If at port of entry the intention is to safeguard against staphylococcal intoxication, a criterion for Staphylococcus aureus would not detect the hazard if the viable cells have died. If epidemiological evidence shows that this product constitutes a hazard, a more direct criterion would then be needed, such as one for thermonuclease or enterotoxin. If a criterion is applied to a finished product it may be inadequate if a destructive process has been applied (e.g., destruction of Staph. aureus, but with enterotoxin persisting). applications. 2nd Ed. International Commission on Microbiological Specifications for Foods.

8 The application and use of criteria 129 The microbiological status of some foods changes continually during storage and distribution and is taken into account when selecting the case. Thus, control at the source is the ideal, and criteria applied at the port of entry will never be as effective, though there are many instances where their application is effective (e.g., Salmonella in dried milk). C. CHOICE OF ORGANISMS The ICMSF has attempted to include in criteria those microbes most important in respect of hazard and/or non-compliance with GCP. This choice was based on epidemiological evidence and expert opinion, recognizing limitations of current methodologies. D. SELECTION OF LIMITS For 3 class attributes plans it was necessary to establish m values (associated with GCP) as well as M values (related to the safety/ quality limit). The latter are based on expert opinion as to the acceptable limit, but the former should be based on firm data obtained from producers and retailers operating according to Good Manufacturing Practice (GMP) and GCP. Although ICMSF has collected a large amount of data, there is not sufficient information for certain commodities to establish m values on this basis. When information was not available, expert opinion was relied upon to establish m values for 3 class plans. Where such values could not be derived, 2 class plans are adopted. Appropriate values of m for 3 class plans must await the collection of further microbiological data. The ICMSF has now implemented a pilot computer-based collection and retrieval system for microbiological data and is anxious to obtain data to expand this data base. Readers having relevant data are invited to contact the secretary of the ICMSF, who will be pleased to provide printed forms for confidentially recording data in a format suitable for the system. REFERENCES ICMSF Microorganisms in Foods. 1. Their Significance and Methods of Enumeration, 2nd ed. University of Toronto Press, Toronto Microbial Ecology of Foods. Vol. 2. Food Commodities. Academic Press, New York applications. 2nd Ed. International Commission on Microbiological Specifications for Foods.

9 11 Sampling plans for raw meats World trade in raw meats, both chilled and frozen, continues to be significant in terms of volume despite their potential hazard to health. Total world red meat production in 1979 was more than 78 million tonnes, with about 9% being traded internationally. A. REASONS FOR SAMPLING Raw meats are important sources of Salmonella and Clostridium perfringens, which are often incriminated in outbreaks of food-borne disease (ICMSF, 1980, p. 354). They are also sources of Staphylococcus aureus, Campylobacterfetus subsp. jejuni, and Yersinia enterocolitica. Salmonellae are found more often in pork and in veal from young calves than in other meats (ICMSF, 1980, p. 347). In the first edition of this book criteria for salmonellae were included as guidelines to assist control of this organism in raw meat. It was hoped that such guidelines would lead to a reduction in the contamination rate and a drop in the incidence of foodborne disease. However, outbreaks of salmonellosis due to meat have continued with little evidence of diminution. In some instances salmonellosis has been caused by the consumption of raw or inadequately cooked meat products but a more common hazard arises through crosscontamination from raw to cooked meat or other foods, and subsequent time-temperature abuse. Clostridium perfringens is ubiquitous, and although it occurs on carcass meat, usually in low numbers, it cannot be controlled by any known means. The majority of outbreaks of C. perfringens gastroenteritis attributable to meat result from inadequate storage of the cooked product. Prevention involves attention to the time/temperature conditions

10 Raw meats 131 of cooking and, more important, to hot holding, cooling, and reheating before consumption. Staphylococcus aureus may occur on raw meat although usually only in low numbers. Contamination by animal strains of Staph. aureus, which are thought to have a low enterotoxin-forming potential, is probably of less consequence than contamination from human sources. Staphylococcus aureus competes poorly with the normal microbial flora of raw meat and constitutes a health hazard only when this competing flora is restricted and there is temperature abuse of the product. Campylobacter fetus subsp. jejuni is often present in the intestinal flora of healthy animals used for food production (Blaser, 1982). However, the numbers present on red meat are generally low and the organism has only a limited potential for growth or survival on refrigerated or cooked meat. Food-borne outbreaks of Campylobacter enteritis appear to arise almost exclusively from eating raw or undercooked food (Blaser, 1982). The use of microbiological criteria as a control measure for reducing the incidence of disease arising from the consumption of meat carrying Salmonella, C. perfringens, and Staph. aureus was considered by a FAO/WHO Working Group (FAO/WHO, 1979) in light of the Codex General Principles for the Establishment and Application of Microbiological Criteria for Foods (Codex, 1981). According to these principles a microbiological criterion should be established and applied only where there is a definite need and where it is both practical and likely to be effective. Such criteria should form part of, or be related to, a Codex Code of Practice for the particular product. The presence of Salmonella and C. perfringens on raw meats is generally more a reflection of their incidence in the live animal than of a breakdown of hygiene. In present circumstances the occurrence of salmonellae and the other pathogens in raw meat cannot be entirely prevented by the application of codes of hygienic practice (ICMSF, 1980, p. 347). Also, the extreme variability of distribution of pathogens such as salmonellae in meats prevents the establishment of practical sampling plans which would ensure with any degree of confidence the absence of salmonellae in meats. Consequently the Working Group concluded that the application of limits for pathogenic microorganisms at port of entry was not appropriate in terms of the Codex General Principles. In view of the importance of Salmonella as a cause of food-borne disease and meat as a vehicle for its transmission there is much to be gained from regular in-plant monitoring to establish the extent of the problem in particular areas and to detect increases in prevalence.

11 132 Proposals for sampling and sampling plans Products that have a history of being implicated as causes of salmonellosis should be sampled at appropriate points during production and distribution to determine the prevalence of contamination and to trace its source on the farm as well as to look at the effects of travel, holding, and processing of stock. The aim is to collect information that will help to motivate the producer to eliminate salmonellae in the live animal and thus control the problem at its source. The ultimate solution to the Salmonella problem lies in producing Salmonella-free animals. The provision of Salmonella-free foodstuffs for animals would be a major advance towards this aim (see Chapter 14, Feeds of Animal Origin and Pet Foods). Until this is achieved treatments that destroy salmonellae in the product (e.g., irradiation) may provide some protection for the consumer. In the absence of effective control measures, environmental and line sampling are more useful than testing the end-products. Although improvement of slaughter hygiene is incapable of eliminating contamination with salmonellae, neglect of hygiene can make the situation much worse. A comprehensive monitoring program in the processing plant can provide a check on the prevalence of infection in livestock and the effectiveness of measures of hygiene in controlling spread of contamination. The FAO/WHO Working Group (FAO/WHO, 1979) concluded also that the number of indicator organisms in meat neither reflects adherence to a code of hygienic practice nor indicates presence or absence of pathogens. Hence criteria based on indicator organisms were not justified for raw meat. These conclusions, and those concerned with pathogens, have been B. SAMPLING PLANS In the virtual absence of studies relating health risks to surface counts of aerobic organisms on carcass meats it is not appropriate to reject consignments of product for public health reasons solely on the basis of high aerobic plate counts. However, aerobic plate counts (APC) can be used to monitor Good Commercial Practice (GCP), and criteria based on such examinations are a valuable aid to in-plant quality control. Microbiological quality control of meat-processing involves development and use of processing methods designed to keep microbial numbers low by reducing contamination and preventing growth. Microbiological monitoring of product and plant at critical control points can be used

12 Raw meats 133 TABLE 19 Sampling plans and recommended microbiological limits for raw meat a Product b Test Method reference c Case Limit per cm 2 or gram Plan class n c m M Carcass meat, before APC chilling Carcass meat, chilled APC Edible offal, chilled APC Carcass meat, frozen APC x Boneless meat, frozen APC x (beef, veal, pork, mutton) Comminuted meat, APC frozen Edible offal, frozen APC x a Not for use at port of entry but for in-plant quality control. Refer to Section B for appropriate applications. b Unfrozen carcasses and primal cuts, swab counts per cm 2 ; other meats and meat products, per gram. c Refers to page number in ICMSF to assess the effectiveness of sanitation factors in limiting microbial growth. These counts are compared with APC criteria established for particular products prepared under specific conditions. Sampling plans and microbiological criteria for certain raw meat commodities are listed in Table 19. The proposals for raw meats are based on limited data collected mainly from production and under a restricted range of circumstances. The criteria have been established as guidelines and some tolerance must be allowed in their application; for example, meat marginally acceptable, or even rejected, by these guidelines could be used in heated products, but would have very limited shelf-life as fresh meat, even under good refrigeration. Meat prepared according to the Recommended International Code of Hygienic Practice for Fresh Meat (CAC/RCP , FAO, Rome) may on occasion exceed these limits, which should not be used as legal standards. No attempt has been made to distinguish between meat from different species although it has been noted that sheep carcasses normally have a slightly higher level of initial

13 134 Proposals for sampling and sampling plans contamination than those of beef and numbers of bacteria tend to be higher again on pig carcasses (ICMSF, 1980, p. 342). Although there are considerable quantities of horse meat in international trade, criteria have not been included because of lack of data. The FAO/WHO Working Group (FAO/WHO, 1979) considered that the only point at which APC could be used for chilled meat, to evaluate the hygienic conditions under which it was produced, was at the slaughterhouse, and because of its perishable nature it would not be possible to set APC limits for use in guidelines or specifications for chilled meats at port of entry. This consideration applies to meat that is vacuum-packaged in an oxygen-impermeable plastic film as well as to chilled meat packaged in other ways. Vacuum-packaging and modified-atmosphere packaging bring about changes in the bacterial flora, and the storage life depends more on the nature of the flora that develops than on the numbers of bacteria present after processing. Case 1 with a 3 class plan for the APC is appropriate for utility tests for general contamination of a product that is normally cooked before consumption. The values for m reflect current commercial attainment based on the examination of meat from different sources, mainly at point of production, and are supported by experience that meat with plate counts lower than the value set for m usually has not been subject to excessive contamination or undue faulty handling. A normal shelf-life would be anticipated. Overall data from a large number of samples have been analysed but not all products have been tested to a similar degree. The M values for APC are based on experience that meat with APC values in excess of 10 7 is either grossly contaminated or has been exposed to conditions permitting microbial growth to a level not far short of the point at which incipient spoilage can be detected. Immediately after slaughter an M value of 10 6 is justified on the basis that counts below this level can be readily achieved under GCP. At this level some tolerance is allowed for further processing which could involve slight increase in microbial numbers. Microbiological criteria applied to frozen meat should be similar to those applied to chilled meat. The criteria proposed for frozen carcass meat and frozen cuts assume that the meat is sampled in the frozen state and thawed quickly before examination of samples. Spoilage occurs at about the same level; hence the M values are the same. Reflecting the small decrease in numbers due to freezing and storage m values are 5 x 10 5 instead of Commercially thawed meat usually has a

14 Raw meats 135 higher count than the frozen product, and the criteria for chilled carcass meat could apply. The microbiological quality of comminuted meat reflects the situation in the meat from which it is prepared. Counts in general are 10 to 100 fold greater in commercial minced meat than on the equivalent carcass (ICMSF, 1980, p. 370). If scrap meat and trimmings from the outer surface of carcasses are used, the counts in the mince will be higher than if whole cuts are minced. Mechanically deboned meat derived from good-quality raw material can be microbiologically equivalent to or better than minced, manually boned meat provided strict temperature control is exercised, but the process needs careful control in respect to the material being boned and sanitation of the equipment. There have been many proposals for microbiological criteria for minced meat but there is no evidence that a criterion has any relevance to health hazard. Accordingly criteria similar to those for carcass meat are proposed. There is little information available on the microbiology of edible offals but the considerations are similar to those for carcass meat and the criteria proposed are the same. C. SAMPLING PROCEDURES (a) Carcasses and primal cuts For chilled or frozen carcass meat and large bone-in or boneless cuts, as n = 5 for all criteria suggested, take subsamples from 5 individual carcasses or cartons of cuts (sample units). If the lot is distributed in several shiploads or freightcars take the sample units from more than one transportation unit. Because contamination of meat is often very uneven, a number of subsamples should be taken from different parts of the carcass or cut, including areas known to be subject to contamination or particularly favourable for microbial growth. The subsamples from a carcass or carton may be pooled and thoroughly mixed to form a composite analytical unit, or each may be treated as a separate analytical unit. Bacterial contamination of carcasses or cuts is generally on the surface, and this should be recognized in the collection of subsamples. Because defacing of carcasses or cuts is often economically undesirable, the surface swab technique is recommended for the APC on unfrozen meat. Samples should be taken at sites likely to be contaminated (see,

15 136 Proposals for sampling and sampling plans e.g., Roberts et al., 1980). Lamb and pork carcasses should be swabbed at at least two points (leg and brisket), and beef and horse at three (leg, flank, and neck). Sampling of cuts should include the original exterior surface and a cut surface. For a comparison of swabbing and excision methods see ICMSF, 1980, p (b) Boneless bulk cuts, comminuted meat, and edible offals Except for the offals it is usually not possible to identify the original surface. From each of 5 cartons or packages remove a sample unit of about 200 g of tissue, which should comprise subsamples from a number of different parts of the pack. This composite sample unit should be mixed thoroughly and the appropriate analytical unit withdrawn. (c) Retail packages These include chilled or frozen consumer-portion packed cuts and offals as well as comminuted meat. The sampling procedure entails the examination of 5 packages from the lot. D. SAMPLE COLLECTION Remove any wrappings from the carcass or package carefully without handling the meat. Use sterile swabs and templates for taking surface swab samples. Mark the sampling area with sterile metal guides (e.g., 5, 50, or 100 cm 2 ). Use two sterile swabs to sample each area. Moisten the first swab with sterile peptone water and rub firmly across the exposed area several times in all directions. Use the second swab dry and rub over the same area. Introduce both swabs into a bottle containing 3 or 4 glass beads and an appropriate known volume of diluent (e.g., 0.1 % peptone, 0.9% NaCl). Shake vigorously. To avoid cross-contamination when removing portions of meat, use sterile instruments (knives for unfrozen meat; saws, cleavers, and special drills for frozen meat), and transfer the piece aseptically to sample containers. Samples of frozen meat should remain frozen during transport to the laboratory. Samples from chilled carcasses should not be frozen.

16 Raw meats 137 E. TEST PROCEDURES (a) Aerobic plate counts Comminute, blend, or stomach the various portions of meat composing the sample unit and withdraw the analytical unit. Combine the swabs to form an analytical unit, or treat each separately. Prepare dilutions and perform the APC test according to the directions given in ICMSF, 1978, p Spread plates are preferable to pour plates when examining raw meats. The temperature used for incubation of plates should be related to the material being examined. To estimate numbers of mesophilic contaminants on freshly dressed carcasses it is advisable to incubate plates at 35 or 37 C. For meat that has been held under chilled conditions, plates should be incubated at 25 C to include psychrotrophs and thus give a better indication of the microbial growth that has occurred. Counts at 20 C give similar results after a longer incubation time. (b) Salmonella The test for salmonellae requires an analytical unit of 25 g and is based on positive or negative results using the method described in ICMSF, 1978, p For frozen products use a non-selective enrichment before selective enrichment and plating on selective agar media. Sample units should be taken from appropriate parts of carcasses, cuts of meat, and offals. REFERENCES Blaser, M.J Campylobacter jejuni and food. Food Technology, 36 (3), Codex. l981. Codex Alimentarius Commission, Fourteenth Session, 1981: Report of the 17th Session of the Codex Committee on Food Hygiene, Washington, D.C November ALINORM 81/13. Appendix ii, General Principles for the Establishment and Application of Microbiological Criteria for Foods. Codex Alimentarius Commission, FAO, Rome FAO/WHO (Food and Agricultural Organization / World Health Organization) Microbiological Criteria for Foods. Report of a Joint FAO/WHO Working Group on Microbiological Criteria for Foods, Geneva, February (Document WG/Microbiol/79/1) WHO, Geneva ICMSF Microorganisms in Foods. 1. Their Significance and Methods of Enumeration, 2nd ed. University of Toronto Press, Toronto

17 138 Proposals for sampling and sampling plans Microbial Ecology of Foods. Vol. 2. Food Commodities. Academic Press, New York Roberts, T.A., MacFie, H.J.H., and Hudson, W.R The effect of incubation temperature and the site of sampling on the assessment of the number of bacteria on red meat carcasses at commercial abattoirs. Journal of Hygiene, Cambridge, 85,

18 12 Sampling plans for processed meats Processed meats comprise a number of meat products, manufactured by many different technologies, including heat-processing, curing, drying, fermenting, acidifying, and packaging (ICMSF, 1980). Processing changes the eating quality of meat. It also changes the composition of the microflora, and some processes kill or prevent growth of pathogens. Basically processing is intended to prolong the shelf-life of the products. The increase in shelf-life may range from a few days to several years. Processed meats with only a slight increase of shelf-life would not be candidates for international trade, while those given some degree of heat-treatment or curing combined with an adequate storage temperature move in international trade. According to the FAO (1981), international trade during 1980 of processed meat and meat preparations amounted to about 1 million tonnes, of which sausages comprised about 115,000 tonnes. These FAO figures relate mainly to three groups of products: (a) shelf-stable cured canned meats; (b) perishable cured canned meats; and (c) cooked or uncooked, fermented, dried, or semi-dried sausages. A. REASONS FOR SAMPLING In the following section processed meats will be treated according to the outline in ICMSF, 1980, pp Discussion will deal with hazards (public health or spoilage), international trade of the products, and the possibility and feasibility of instituting means of control other than by microbiological criteria.

19 140 Proposals for sampling and sampling plans TABLE 20 Sampling plans and recommended microbiological limits for processed meats a Product Test Method reference b Case Limit per Plan gram class n c m M Dried blood, plasma, and gelatin Staph. aureus C. perfringens Salmonella c Roast beef d Salmonella 163 c Pâté Salmonella 163 c a Refer to text for appropriate applications. b Refers to page numbers in ICMSF, c See also ISO 6579 (1981). d Includes beef cooked in water baths; see Section B(f). B. SAMPLING PLANS (a) Dried meats and dried animal products Dried meats like jerky, charqui, and biltong are mainly produced and consumed locally. Most dried meats found in international trade are intended for further processing. Further processing would normally consist of dry-blending with other food ingredients (e.g., dried soups) or rehydration and canning. In both cases the dried meats are subjected to conditions that either would not change or would reduce the hazards. In international trade no evidence of hazard has been demonstrated, and no microbiological criteria are proposed. Dried animal products such as dried whole blood, blood plasma, and gelatin are used as ingredients for formulated foods. When these ingredients are used in further processed products that are not subjected to adequate heat-treatment it is proposed to include sampling plans for Staphylococcus aureus, Clostridium perfringens, and Salmonella (Table 20). For Staph. aureus and C. perfringens 3 class sampling plans, case 8 with n = 5, c = 1, m = 10 2, M = 10 4, are proposed. Salmonella should be considered in case 11 with a 2 class sampling plan with n = 10, c = 0.

20 Processed meats 141 (b) Raw cured meats, high a w High a w meat products are defined as those with an a w above 0.92, such as occurs in bacon produced by a Wiltshire or similar process (ICMSF, 1980, pp ). In Wiltshire curing sides or parts of sides of pork, with or without bones, are injected with brine before either tank- or dry-curing. Smoke may be applied, but the product is not heated. There are many other types of manufacture of bacon, but this is the most common for bacon in international trade. The microbiology of bacon is characterized by a rapid change from the initial gram-negative flora to gram-positive salt-tolerant micrococci and lactic acid bacteria. Only where curing is inadequate or the ph of the meat (usually the collar) is high will putrefactive spoilage dominate. Otherwise, spoilage is characterized by sliming. Staphylococci could be considered to be of concern but are not able to compete with the normal saprophytic flora. Applying microbiological criteria at manufacture can make little contribution to the control of spoilage. Salmonella is not a major concern because the product is cooked immediately before consumption. (c) Raw, cured, or salted meats and natural casings, low a w Low a w meat products are defined (ICMSF, 1980) as those having a water activity of 0.92 or below. This group includes whole cuts of meat (e.g., Bundener Fleisch, Parma ham, Westphalian ham, Smithfield ham, Prosciutti), non-fermented dried sausages, and natural casings, all of which are in international commerce. Although some of the raw cured or salted meats have been implicated in outbreaks of food-borne illness, particularly botulism, the incidents have usually involved home-prepared products. Commercial products have had a very good record of safety. Growth of nonpathogenic bacteria occurs during the process of salting, curing, and drying of many of these products and can contribute to their flavour characteristics. Hence, microbiological criteria need not be applied to these commercial products. Cured, dried non-fermented sausages are in international trade but the only spoilage problem appears to be mould formation on the casing. No public hazards have been identified, and no microbiological criteria are proposed. Natural casings from international commerce are generally packed

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